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Poly-β-hydroxybutyrate (PHB) Depolymerase fromFusarium solaniThom
- Source :
- Journal of Chemistry, Vol 2013 (2013)
- Publication Year :
- 2013
- Publisher :
- Hindawi Limited, 2013.
-
Abstract
- Fusarium solaniThom produced maximum PHB depolymerase by 48 h when grown in BHM containing 0.2%, w/v PHB, pH 8.0 at25°C. Statistical optimization studies using Plackett Burman design of PHB depolymerase production yielded maximum PHB depolymerase activity after 2 days as against 4 days in the unoptimized conditions with a 2-fold increase in activity. Partial purification of the extracellular poly-β-hydroxybutyrate (PHB) depolymerase PHAZFusfromF. solaniThom by two steps using ammonium sulphate (80% saturation) and affinity chromatography using concanavalin-A yielded 162.3-fold purity and 63% recovery of protein. The enzyme composed of a single polypeptide chain of 85 KDa, as determined by SDS-PAGE. The enzyme stained positive for glycoprotein by PAS staining. Optimum enzyme activity was detected at pH 7.0 and55°C. The enzyme was stable at pH 7.0 and 55°C for 24 h with a residual activity of almost 85%. Paper chromatography revealedβ-hydroxybutyrate monomer as the major end product of PHB hydrolysis. Complete inhibition of the enzyme by 1 mM HgCl2(100%) indicated the importance of essential disulfide bonds (cystine residues) for enzyme activity or probably for maintaining the native enzyme structure.
- Subjects :
- chemistry.chemical_classification
Chromatography
Article Subject
biology
Plackett–Burman design
Chemistry
General Chemistry
biology.organism_classification
Enzyme structure
Enzyme assay
lcsh:Chemistry
Paper chromatography
Hydrolysis
Enzyme
lcsh:QD1-999
Biochemistry
Affinity chromatography
biology.protein
Fusarium solani
Subjects
Details
- ISSN :
- 20909071 and 20909063
- Volume :
- 2013
- Database :
- OpenAIRE
- Journal :
- Journal of Chemistry
- Accession number :
- edsair.doi.dedup.....7fdc75975ea525637c8efbee1aae743e